Am. J. Respir. Cell Mol. Biol., Volume 18, Number 3, March, 1998 353-362

Organization of the Human SP-A and SP-D Loci at 10q22-q23
Physical and Radiation Hybrid Mapping Reveal Gene Order and Orientation

Russell R. Hoover and Joanna Floros

Departments of Cellular and Molecular Physiology and Pediatrics, Pennsylvania State University College of Medicine, Hershey, Pennsylvania

The human surfactant protein (SP) A locus has been assigned to chromosome 10q22-q23 and consists of two very similar genes, SP-A1 and SP-A2, as well as a truncated pseudogene. SP-A belongs to the family of collagenous C-type lectins along with mannose binding protein (MBP) and SP-D, both of which have also been mapped to the long arm of chromosome 10. In this article we report the relative location and orientation of each of the SP-A and SP-D genomic sequences. Characterization of two overlapping genomic clones revealed that the SP-A pseudogene lies in a reverse orientation 15 kb away from the 5' side of SP-A1. This finding was verified by the amplification of the entire SP-A pseudogene/SP-A1 intergenic region using long-range polymerase chain reaction. The relative location of SP-A2 and SP-D was then ascertained by testing a number of sequence tagged sites against the Stanford TNG3 and G3 radiation hybrid panels. The radiation hybrid mapping data showed that both SP-A2 and SP-D are on the 5' side of SP-A1 at approximate distances of 40 kb and 120 kb, respectively. The SP-A and SP-D loci were also oriented relative to the centromere, with the overall order being: centromereSP-DSP-A2pseudogeneSP-A1 telomere.

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